The novel CRISPR/Cas9 has been one of the most progressive advancements of the molecular biology world giving us the tool to edit genomes. The way CRISPR/Cas 9 works is by first, cleaving DNA at a specified point guided by Cas9 then second, repairing the cleavages of the DNA with by non-homologous end joining (NHEJ). This tool has proven to be helpful in the field of virology especially in engineering large DNA viruses. One such virus is the Vaccinia virus known for its use in the eradication of smallpox.
Vaccinia virus is a large double-stranded DNA virus with flexibility of accommodating insertions and deletions to its genome. Vaccinia virus is a good candidate for genome editing by homologous recombination due to its ability to carry its own repair machinery. In this paper, investigators show that the genome cutting of Vaccinia virus using CRISPR/Cas9 mechanism is efficient but not the NHEJ repair resulting in the inhibition of replication of the genome. This was shown in the suppressed spread of the virus in culture. However, the investigators show that this outcome indicates that CRISPR/Cas 9 can actually be used as a powerful tool for selecting poxvirus recombinants which would otherwise be impossible to separate without gene markers. Through this method of selecting for recombinant Vaccinia virus, the generation of poxvirus-based vaccines can be sped up - to under 2 weeks-, proving CRISPR/Cas9 to be an attractive platform for personalized cancer vaccines and emerging poxvirus disease outbreak control.
No comments:
Post a Comment